This proposal seeks to continue and expand technological biomedical research, collaborative, service, training and dissemination activities of the Laboratory for Fluorescence Dynamics (LFD). Since 1986, the LFD has been a national resource center dedicated to 1) developing new fluorescence technologies for biology and medicine and 2) service, training, and dissemination of fluorescence methods in a user facility. In previous years (1986-2010), we established our reputation as the leaders in technological development for fluorescence dynamics. Since moving to UCl in 2006, we expanded the core research and collaborative work in this new fertile biomedical research environment. We created structures for training through workshops, tutorials and organization of LFD training courses at UCl and other US universities. Core development expands current limits of biomedically relevant fluorescence instrumentation, particularly for the study of cellular processes. Projects include: 1) new concepts in fluorescence microscopy that develop novel imaging methods to exploit the dynamic fluorescence methodologies that are the hallmark of the LFD developments, 2) expanding the concept of spatio-temporal fluctuation correlation analysis to a new powerful image approach that provides detailed information of molecular flow at the nanoscale, 3) new methods for 3D nanoimaging based on the orbital tracking approach, 4) novel technologies for fluorescence lifetime imaging analysis that allow new contrast mechanisms to be applied to tissues and animals, and 5) development of software/algorithm platforms for disseminating the methods for data collection and analysis developed at the LFD. Users at the LFD find ready access to new and innovative technology, which affords them a unique opportunity to rapidly advance their own research programs. Driving biological problems exploit and push technological advances and research in biological processes, macromolecular assembly, tissue organization and dynamics, membrane morphology/function relationships, and innovative biomedical instrumentation. Educational programs include hands-on training of students, postdoctoral fellows and visiting scientists, workshops and specialized schools in innovative fluorescence methodologies.